Proliferation conditions for human satellite cells. The fractional content of satellite cells.
APMIS
; 109(11): 726-34, 2001 Nov.
Article
en En
| MEDLINE
| ID: mdl-11900051
Primary satellite cell cultures have become an important tool as a model system for skeletal muscles. A common problem in human satellite cell culturing is fibroblast overgrowth. We combined N-CAM (Leu19) immunocytochemical staining of satellite cells (Sc) with stereological methods to estimate the fraction of Sc in culture. Evaluation of different culture conditions allowed us to find proliferation conditions preferentially for Sc: a) Sc should be cultured on surfaces coated with ECM-gel. b) Primary cell culture should be inoculated in DMEM supplemented with 10% fetal calf serum to increase cell adherence. c) Change of media to DMEM supplemented with 2% Ultroser-G and 2% FCS after 24 h.d) Before subcultivation, cells should be preplated for 30 min. The fractional content of Sc in passage four when applying this method of cultivation was 0.82 +/- 0.07 (mean +/- SE, N = 10). Our method enabled us to establish culture conditions which resulted in high Sc content despite several subcultivations. Estimation of the fractional cell content could be a useful tool for optimizing not only Sc-culturing but all cultures initially containing more cell types.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Células Madre
/
Músculo Esquelético
Límite:
Humans
Idioma:
En
Revista:
APMIS
Asunto de la revista:
ALERGIA E IMUNOLOGIA
/
MICROBIOLOGIA
/
PATOLOGIA
Año:
2001
Tipo del documento:
Article
País de afiliación:
Dinamarca
Pais de publicación:
Dinamarca